Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for isolating and culturing eyebag adipose-derived stem cell in vitro

A technology for adipose stem cells and in vitro culture, which is applied in the field of eye bag adipose stem cell separation and in vitro culture, which can solve the problems of low separation efficiency, cumbersome operation steps, and increased risk of cell damage, and achieve reduced cell damage, good viability, and good cell condition Effect

Inactive Publication Date: 2019-01-25
希瑞干细胞科技有限公司
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, most of the separation methods of adipose stem cells are separated by enzymatic digestion after mechanical treatment. The operation steps are cumbersome and the separation efficiency is low. In the later stage of culture, more double antibodies and serum are added. Although it has certain benefits for the culture, it is also Increase the damage to cells and the risk of later clinical use

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for isolating and culturing eyebag adipose-derived stem cell in vitro
  • Method for isolating and culturing eyebag adipose-derived stem cell in vitro
  • Method for isolating and culturing eyebag adipose-derived stem cell in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for the separation and in vitro culture of eye bag fat stem cells proposed by the present invention comprises the following steps:

[0024] 1. Reagent preparation:

[0025] 1) Preparation of cleaning solution: mix physiological saline and double antibody according to the volume ratio of 500:5;

[0026] 2) Preparation of digestive enzyme solution: 0.1% collagenase type I, 0.05% collagenase type IV, and 0.25% trypsin / EDTA are prepared by mixing at a volume ratio of 1.5:3:1;

[0027] 3) Preparation of serum-free medium: Lonza serum-free medium, PALL serum substitute, and 20mmol / L L-glutamine solution were prepared by mixing at a volume ratio of 500:10:5.

[0028] 2. Acquisition of pouch fat tissue: Pouch fat tissue is the fat waste that women and children remove from eye pouch surgery of the elderly, and is stored in sterile normal saline at 4°C for 4 hours.

[0029] 3. Separation of eye bag fat stem cells: Rinse the eye bag fat tissue 3-5 times with cleaning so...

Embodiment 2

[0032] A method for the separation and in vitro culture of eye bag fat stem cells proposed by the present invention comprises the following steps:

[0033] 1. Reagent preparation:

[0034]1) Preparation of cleaning solution: mix physiological saline and double antibody according to the volume ratio of 500:5;

[0035] 2) Preparation of digestive enzyme solution: 0.1% type I collagenase, 0.05% type IV collagenase, 0.25% trypsin / EDTA mixed at a volume ratio of 2:2:1;

[0036] 3) Preparation of serum-free medium: Lonza serum-free medium, PALL serum substitute, and 20mmol / L L-glutamine solution were prepared by mixing at a volume ratio of 500:10:5.

[0037] 2. Acquisition of pouch fat tissue: Pouch fat tissue is the fat waste that women and children remove from eye pouch surgery of the elderly, and is stored in sterile normal saline at 4°C for 4 hours.

[0038] 3. Isolation of eye bag adipose stem cells: wash the eye bag fat tissue with cleaning solution for 3 times, remove the b...

Embodiment 3

[0052] A method for the separation and in vitro culture of eye bag fat stem cells proposed by the present invention comprises the following steps:

[0053] 1. Reagent preparation:

[0054] 1) Preparation of cleaning solution: mix physiological saline and double antibody according to the volume ratio of 500:5;

[0055] 2) Preparation of digestive enzyme solution: 0.1% type I collagenase, 0.05% type IV collagenase, 0.25% trypsin / EDTA are prepared by mixing at a volume ratio of 3:1.5:1;

[0056] 3) Preparation of serum-free medium: Lonza serum-free medium, PALL serum substitute, and 20mmol / L L-glutamine solution were prepared by mixing at a volume ratio of 500:10:5.

[0057] 2. Acquisition of pouch adipose tissue: Pouch adipose tissue is fat waste removed from women and children from elderly pouch operations, and stored in sterile normal saline at 4°C for 4 hours.

[0058] 3. Isolation of eye bag adipose stem cells: wash the eye bag fat tissue with cleaning solution for 3 times...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

A method for isolating and culturing eyebag adipose-derived stem cell in vitro is characterized by comprising that following step: obtaining adipose tissue, isolating adipose-derived stem cells and culturing adipose-derived stem cells in vitro. The invention has the advantages that compared with the prior art, the method for isolating and culturing the adipose stem cells in vitro has the followingadvantages: 1, the cells are cultured in the serum-free culture medium without the addition of the heterologous serum components, thereby reducing the injury to the cells and the risk of the later clinical use; 2, that cell state is good, the viability is good, the cell shape, the proliferation ability, the surface mark expression and the differentiation ability are good.

Description

technical field [0001] The invention relates to the technical field of stem cell separation and in vitro culture, in particular to a method for the separation and in vitro culture of eye bag fat stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are the hotspot cells of adult stem cells with multi-lineage differentiation potential. , Extracted from fat, under specific culture conditions, mesenchymal stem cells can differentiate into fat cells, osteoblasts, chondrocytes and muscle cells. Mesenchymal stem cells are currently the main seed cells for tissue engineering. After many years of research by many scholars, although tissue engineering has made great progress, however, due to the lack of ideal screening methods, the difficulty of obtaining mesenchymal stem cells, and the difficulty of in vitro expansion, it has become a viable option for reinfusion at the clinical level. Application is limited. Therefore, it is one of the urgent problems to be sol...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0667C12N2500/32C12N2500/90C12N2509/00
Inventor 费方利王浩
Owner 希瑞干细胞科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com